Development and evaluation of an enzyme-linked immunosorbent assay based on a recombinant SpaA protein (rSpaA415) for detection of anti-Erysipelothrix spp. IgG antibodies in pigs

L G Giménez-Lirola, C T Xiao, P G Halbur, T Opriessnig

Research output: Contribution to journalArticlepeer-review

Abstract

The aim of this study was to develop an enzyme-linked immunosorbent assay (ELISA) for detection of anti-Erysipelothrix spp. IgG in pig sera by utilizing recombinant polypeptide SpaA415 (rSpaA415) based on surface protective antigen (Spa) A (SpaA) of Erysipelothrix spp. The sensitivity of the rSpaA415 ELISA was evaluated on sera from pigs experimentally infected with E. rhusiopathiae serotype 1a (n=72), serotype 19 (n=12), or experimentally vaccinated with a commercial attenuated-live vaccine based on serotype 1a (n=12), a commercial bacterin based on serotype 2 (n=12), or an experimental bacterin based on serotype 2 (n=300). Specificity was tested using 221 negative control samples. The earliest antibody response was detected at 7 days post-inoculation (dpi) and 14 days post-vaccination (dpv). At the cutoff of 0.9 sample optical density, the sensitivity was 96.5% and the specificity was 100%. In experimentally infected pigs, the sensitivity of the rSpaA415 ELISA ranged from 5.5 to 100% which improved as dpi increased. Antimicrobial treatment, administered prior to appearance of clinical signs, decreased assay sensitivity. In vaccinated pigs, the rSpaA415 ELISA had a sensitivity of 48.3-100%. Serum samples from rabbits each hyperimmunized with one of the 28 Erysipelothrix spp. serotypes were used to determine cross-reactivity with strains expressing SpaB, SpaC or no currently recognized Spa protein and antibodies against E. tonsillarum were not detected. These data suggest that the novel rSpaA415 ELISA test is a useful tool to detect anti-IgG antibodies against different serotypes of E. rhusiopathiae in infected or vaccinated pigs without cross-reacting with the economically less important E. tonsillarum strains.
Original languageEnglish
Pages (from-to)191-7
Number of pages7
JournalJournal of microbiological methods
Volume91
Issue number1
DOIs
Publication statusPublished - Oct 2012

Keywords

  • Animals
  • Antibodies, Bacterial
  • Antigens, Bacterial
  • Clinical Laboratory Techniques
  • Cross Reactions
  • Enzyme-Linked Immunosorbent Assay
  • Erysipelothrix
  • Erysipelothrix Infections
  • Immunoglobulin G
  • Recombinant Proteins
  • Sensitivity and Specificity
  • Swine
  • Swine Diseases
  • Veterinary Medicine

Fingerprint

Dive into the research topics of 'Development and evaluation of an enzyme-linked immunosorbent assay based on a recombinant SpaA protein (rSpaA415) for detection of anti-Erysipelothrix spp. IgG antibodies in pigs'. Together they form a unique fingerprint.

Cite this